The goal of our project is to create an advanced antibacterial first aid kit by combining multiple material binding domains with various antimicrobial peptides. Our Parts are categorized into two main sections: the antibacterial functional domain and the material binding domain.

The first section focuses on the antibacterial functional domain (Table 1), where we characterized four defensins (HNP1, HNP4, HD5, HBD3) along with four HNP1 mutants (HNP1Ala, HNP1ANF, HNP1AWW, HNP1AWK). HNP1 and HNP4 are newly uploaded parts that have been functionally verified, while HD5 and HBD3 are updated parts with confirmed functions. The newly designed defensins, HNP1Ala, HNP1AWW, and HNP1AWK, exhibit antibacterial activity but show reduced effectiveness against Staphylococcus aureus compared to HNP1; HNP1ANF was not expressed.

Table 1: Part Collection of Defensins

The second section is the material binding domain. We characterized six binding domains, including cellulose-binding domains CBM2 and CBM3. We updated the function of CBM3 and confirmed the functionality of CBM2 for the first time. Additionally, CBM5, α1, α2, and vbCBMxx serve as binding domains for chitosan, collagen, and alginate, respectively, and their functions were also newly verified.

Table 2: Part Collection of Binding Domains

After assembling these two groups of parts, we combined them to assess their antibacterial capabilities. The combinations of CBM3 with the four defensins and HNP1 with three binding domains demonstrated antibacterial functions. Unfortunately, the combination of CBMxx and HNP1 could not be successfully purified due to bacterial contamination. Nevertheless, these results indicate that the material binding and antibacterial functional domains we collected can be freely combined, achieving antibacterial properties through the use of the rtUlp enzyme cleavage we characterized, thus accommodating a variety of application scenarios.

Table 3: Part Collection of Binding Domain-sumo-Defensins and Sumo Protease